Monoclonal antibody fragment from combinatorial phage display library neutralizes alpha-latrotoxin activity and abolishes black widow spider venom lethality, in mice

Toxicon : Official Journal of the International Society on Toxinology
Francesca BugliM Pescatori

Abstract

Alpha-latrotoxin (alpha-ltx), a component of the venom of black widow spiders (BWSV), binds to higher vertebrates presynaptic nerve terminals, stimulating massive neurotransmitter release. This neurotoxic protein is responsible for most of the symptoms elicited in men by the bite of black widow spider (BWS), i.e. a neurological syndrome named latrodectism. By reasoning that targeting this single component would abrogate most of the effect of BWS envenomation, we took advantage of the antibody phage display technology to generate monoclonal Fab fragments able to bind and neutralize the alpha-ltx. To this aim, we immunized Balb/c mice with purified toxin and cloned their antibody repertoire in the pCombIII phage display vector. By combining a high-stringency affinity selection with a sensitive 45Ca(2+) uptake assay, we isolated a Fab fragment (FM1) able to bind the alpha-ltx in the low nM range and neutralize its ionophore activity, in vitro and in vivo. After the onset of overt symptomatology, administration of FM1 to experimentally envenomed mice induced remission of symptoms and prevented lethality. Since alpha-ltx is the only molecule responsible for the great toxicity of BWS bites in mammals, the FM1 Fab, highly effective in...Continue Reading

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Citations

Jun 18, 2014·International Journal of Nanomedicine·Francesca BugliAlessandro Arcovito
Jul 24, 2013·Clinical & Developmental Immunology·Francesca BugliFrancesco Paroni Sterbini
May 22, 2009·Journal of Proteomics·G P Espino-SolisL D Possani
Nov 17, 2012·BMC Biotechnology·Rajesh KumarKalpana Luthra
Oct 18, 2011·Biochemical Pharmacology·Andis GraudinsGraham M Nicholson
Oct 8, 2011·Toxicon : Official Journal of the International Society on Toxinology·V Quintero-HernándezL D Possani
Jul 28, 2016·Toxins·Andreas Hougaard LaustsenMikael Engmark
May 21, 2016·Proteomics. Clinical Applications·Philipp KuhnMichael Hust
Jul 30, 2019·Frontiers in Immunology·Manuela B PuccaAndreas H Laustsen
May 24, 2020·International Journal of Molecular Sciences·Steven Ascoët, Michel De Waard
Jan 20, 2021·Toxicon : Official Journal of the International Society on Toxinology·Ana Luísa Soares de MirandaBenito Soto-Blanco

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